A convenient and sensitive fluorescence assay for phospholipid vesicles using diphenylhexatriene |
| |
Authors: | E London GW Feigenson |
| |
Institution: | Section of Biochemistry, Molecular and Cell Biology, Clark Hall, Cornell University, Ithaca, New York 14853 USA |
| |
Abstract: | When phospholipid vesicles are added to an aqueous solution of 1,6-diphenyl-1,3,5-hexatriene (DPH) a fluorescence enhancement of up to several hundredfold is observed which can be used for a determination of phospholipid concentration. Fluorescence enhancement of 2 μm DPH is proportional to the phospholipid concentration over a wide range. As little as 0.7 nmol (~0.5 μg of phospholipid) can be determined to within ±10%. The fluorescence is a function of the type of phospholipid used, salt concentration, and time of incubation. Protein and detergents also enhance DPH fluorescence but to a much smaller extent. Optimal conditions for the assay are presented. Use of this assay to detect phospholipid vesicles fractionated by size on a Sepharose 4B column is illustrated. In this case the method compares favorably to more classical methods of analysis in terms of sensitivity, accuracy, and time involved. |
| |
Keywords: | Authors to whom all correspondence should be addressed |
本文献已被 ScienceDirect 等数据库收录! |
|